The goal of this project is to develop a new highly sensitive and cost effective RNA Bioluminescent Assay (RT-qBLA) for high-throughput detection and quantification of RNA and DNA in biological samples. The assay implements the same detection concept known from pyrosequencing, yet expanding pyrosequencing methodology for highly sensitive and accurate quantification of RNA and DNA molecules. The proposed assay has unique sensitivity and dynamic range and is able to measure kinetics of RNA and DNA replication in the solution in real time without long and expensive sample amplification of the real time PCR. The RT-qBLA requires less expensive reagents and equipment than real time PCR and DNA microarrays and can be used in a various commercial assays in life sciences research, drug discovery, and clinical diagnosis.